Abstract
5515^ Background: Predicting individual responses of HNSCC to treatment remain challenging. Better methods of defining HNSCC are needed. Improvements in biomarker research will probably be achieved with sets of various genomic and proteomic markers as provided by microarray technology. In previous work we demonstrated that the pattern of plasma exosomal-miRNA mirrored the miRNA within the tissue. Here we report the potential for blood-borne miRNA to identify HNSCC and to predict outcome. Methods: Plasma samples (1ml) were obtained from patients diagnosed with HNSCC at the time of initial treatment and serial samples were also obtained during clinical follow-up. Circulating exosomes were isolated and total RNA was extracted by a modified Trizol protocol. Small RNA was subsequently isolated using a small RNA isolation kit (SABiosciences). The specific miRNAs within the small RNA were quantitated using a cancer specific qRT-PCR array (SABiosciences) with an Agilent M3005P. The miRNA expression was compared to those expressed in lung cancer on a prior work. Results: Thirty HNSCC patients (19M, 11F), stage II (n=3), III (n=7), IVa (n=20) entered in the study. All but one underwent radiation and a radiosensitizing agent. Fifteen were considered for analysis since at least one follow-up sample was collected post treatment. Patients with HNSCC exhibited miRNA profiles within circulating exosomes that were distinct from normal controls and patients with squamous cell carcinoma (SCC) of the lung. Of the 84 miRNAs analyzed and 12 detected, miR148b and miR222 appeared to be uniquely expressed in HNSCC. miR16 was present in exosomes on patients with SCC. Patients with lung SCC appear to uniquely express miR199a and miR200c. In patients responding to initial therapy, the levels of most exosomal-miRNAs were suppressed; in patients failing to respond, no decrease in the exosomal-miRNAs were observed. Conclusions: miRNA profiles within blood-borne exosomes may have utility in the HNSCC diagnosis, and appear to be useful as disease monitoring. While these findings need to be confirmed in larger studies, determination of miRNA profiles within plasma-derived exosomes will add a new tool to histology directed therapy.
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